Confined proteins (in synthetic matrices, e.g. sol-gels) as a class of materials have significant potential utility for a broad range of biomedical and biotechnology applications. In most situations, long-term protein stability with either retention or enhancement of activity is required. To fully harness the biomedical implications of confined proteins, it is essential to understand the biophysical mechanisms of how confinement and co-solutes (i.e. osmolytes) modulate the relevant protein properties. The proposed project is built on the growing realization that protein dynamics determine stability and reactivity and that these functionally important protein dynamics are in turn significantly modulated by hydration shell waters. The proposed project will use a combination of spectroscopic, kinetic and simulation based approaches to determine on a molecular biophysics level, how confinement in the presence and absence of osmolytes alters hydration shell water interactions and how these alterations in hydration shell properties impact functionally important protein dynamics. This overall objective will be pursued through two specific aims: Specific aims 1. Determine how confinement in the presence and absence of added osmolytes modifies hydration shell waters of select proteins and peptides. Specific aims 2. Determine how confinement in the presence and absence of added osmolytes impacts functionally important categories of protein dynamics. .